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COI sequences were manually aligned and 12S and 16S were automatically aligned using ClustalW multiple alignment implemented in BioEdit version 7.0.5.3 [22].
The database alignment implemented in Sybyl7.3 [16] was used to align 102 compounds using most active compound 32 as a template.
Another useful feature of the MG-CFA with alignment implemented in Mplus is the possibility of calculating the statistical significance of the difference between the latent means.
The branch robustness of ML trees was estimated using the non parametric bootstrap procedure (100 replicates of the original alignment) implemented in Treefinder with the same parameters than for ML tree inference.
Ambiguous alignment of the tRNA and rRNA genes were discarded from the analyses based on the 90% minimum posterior probability of correct alignment implemented in ProAlign, leaving 938 and 1474 nucleotide characters from 22 tRNA and 12S+16S rRNA genes, respectively.
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Counts of changes per synonymous site were calculated between all sequence pairs in each gene family, based on in-frame codon alignments, implemented in the SNAP program [55].
Phylogenetic analyses using maximum likelihood were performed on the Gblocks-reduced amino acid sequence alignments implemented in the program RAxML 7.2.8 [ 67].
Also sequences with regions of unspliced introns were corrected and aligned using MUSCLE alignment tool implemented in Molecular Evolutionary Genetic Analysis software (MEGA 5.0).
These sequences, together with the nuclear, nucleomorph, plastid, and cyanobacterial sequences were aligned using the progressive alignment procedure implemented in CLUSTALX [ 29], edited to remove any ambiguously aligned regions, and conserved blocks of aligned residues (containing 495 amino acids) were used for phylogenetic analyses.
Phylogenetic analyses for the matR gene were performed at the nucleotide (codon) and the amino acid level, and for the 18S gene at the nucleotide level, using ML and full ML bootstrapping (100 replicates), with the GTR + GAMMA substitution model for the nucleotide alignment and the JTT model for the AA alignment, as implemented in RAxML [ 48].
Among the various algorithms for aligning multiple sequences, we selected the alignment approach implemented in the PRANK program [16].
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